CN113999244A - 4H-pyrano [2,3-c ] pyridine-4-ketone compound and preparation method thereof - Google Patents

Abstract

The invention relates to the technical field of medicaments, and provides 4H-pyrano [2,3-c ]]Pyridine-4-ketone compounds and a preparation method thereof. The 4H-pyrano [2,3-c ]]The pyridine-4-ketone compound is a compound with a structure shown in a formula I or a pharmaceutically acceptable salt thereof. The structure of the formula I is as follows:

wherein R is₁Is R₁Is substituted or unsubstituted phenyl or arylheterocyclyl, R₂Is a 1H-indazol-6-yl group,Benzo [ d ] carbonyl][1,3]Dioxolan-5-yl. The 4H-pyrano [2,3-c ]]Pyridin-4-ones are useful as effective PLK1 inhibitors. The pharmaceutical composition containing the compound also has good anti-tumor physical activity.

Description

4H-pyrano [2,3-c ] pyridine-4-ketone compound and preparation method thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a 4H-pyrano [2,3-c ] pyridine-4-ketone compound and a preparation method thereof.

Background

Polo-like kinases (PLKs) are a class of serine/threonine protein kinases with highly conserved structures and functions, are important regulators of the cell cycle, and can participate in the regulation of different stages of the cell cycle. The four reported subtypes of PLKs (PLK 1, PLK2, PLK3, PLK 4) are structurally similar, with the highly conserved ATP domain at the N-terminus and the characteristic PBD domain for catalytic activity and subcellular dynamic localization at the C-terminus. Among them, PLK1 is involved in the regulation of cell division, DNA damage repair pathways, apoptosis and key steps in cell cycle progression, plays an important role in multiple steps during the process from the beginning of the M phase of mitosis to the end of mitosis, and mainly regulates G2/M phase action on B1 and CDK1 proteins of cells, and in mitosis, mucin is removed from sister chromatid arms, activating APC/C ubiquitin E3 ligase complex, thereby affecting spindle formation, chromosome assembly and separation, cytokinesis, microtubule and centromere interaction in mitosis.

Over-expression of PLK1 promoted cell proliferation, increased centrosome instability and heteroploidy leading to tumor formation. Research shows that the PLK1 inhibitor can effectively inhibit the proliferation of tumor cells. Various PLK1 inhibitors are currently in phase i/ii clinical trials. Therefore, the development of the PLK1 inhibitor with novel structure and better activity is of great significance.

Disclosure of Invention

In order to search for a novel PLK1 inhibitor, the invention designs and synthesizes a series of 4H-pyrano [2,3-c ] pyridine-4-ketone compounds which have novel structures and high inhibitory activity on Polo-like kinase through extensive and intensive research, and the inhibitory activity of Polo-like kinase of the compounds is researched.

An object of the present invention is to provide a 4H-pyrano [2,3-c ] pyridin-4-one compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a process for the preparation of the above compound.

It is another object of the present invention to provide a pharmaceutical composition comprising the above compound.

Another object of the present invention is to provide the use of the above compounds and the above pharmaceutical compositions as PLK1 inhibitors for the treatment of tumors.

In order to achieve the purpose, the invention adopts the technical scheme that:

in a first aspect, the present invention provides a compound having the structure of formula i:

wherein R is₁Is a substituted or unsubstituted phenyl or aromatic heterocyclic group,

R₂is 1H-indazol-6-yl, benzo [ d][1,3]Dioxolan-5-yl.

Further, the aromatic heterocyclic ring is a six-membered aromatic heterocyclic ring. Preferably, the aromatic heterocycle is pyridine.

Further, said R₁Is phenyl or an aromatic heterocyclic group substituted by at least one group selected from cyano, amido, fluorine, chlorine, bromine and methoxy.

Further, the compound having the structure of formula i or a pharmaceutically acceptable salt thereof provided by the present invention may be selected from compounds having the following structure:

in a second aspect, the present invention provides a process for the preparation of a compound having the structure of formula i or a pharmaceutically acceptable salt thereof, comprising the steps of:

and (3) synthesis of an intermediate III:

reacting a compound II with a structure shown in a formula II and ethyl formate in a first solvent in the presence of a first base at a first reaction temperature to generate an intermediate III with a structure shown in a formula III;

and (3) synthesizing an intermediate IV:

the intermediate III reacts in a second solvent under the action of a bromization reagent at a second reaction temperature to generate an intermediate IV with a structure shown in a formula IV;

synthesis of intermediate vi:

under the action of a catalyst, the intermediate IV and a compound V with a structure of a formula V are subjected to a coupling reaction in a third solvent in the presence of a second alkali at a third reaction temperature to generate an intermediate VI with a structure of a formula VI;

and (3) synthesizing an intermediate VII:

the intermediate VI reacts in a fourth solvent under the action of phosphorus oxychloride at a fourth reaction temperature to generate an intermediate VII with a structure shown in a formula VII;

synthesis of Compound I:

and reacting the intermediate VII and the compound VIII with the structure of the formula VIII in a fifth solvent in the presence of a third base at a fifth reaction temperature to generate the compound with the structure of the formula I.

As a preferable embodiment of the preparation method provided by the present invention, the first solvent is at least one of tetrahydrofuran, dioxane, toluene, and acetonitrile;

and/or the second solvent is at least one of tetrahydrofuran, dioxane, N-dimethylformamide and acetonitrile;

and/or the third solvent is any one of dioxane, N-dimethylformamide, toluene, N-dimethylacetamide, acetonitrile and dioxane/water;

and/or the fourth solvent is at least one of acetonitrile, toluene and N, N-dimethylformamide;

and/or the fifth solvent is at least one of N, N-dimethylacetamide, N-dimethylformamide, toluene, acetonitrile and dimethyl sulfoxide.

As another preferable embodiment of the preparation method provided by the present invention, the first reaction temperature is 0 to 120 ℃;

and/or the second reaction temperature is 0-80 ℃;

and/or the third reaction temperature is 40-120 ℃;

and/or the fourth reaction temperature is 20-120 ℃;

and/or the fifth reaction temperature is 20-120 ℃.

As another preferable embodiment of the preparation method provided by the present invention, the first base is at least one of sodium hydride, sodium tert-butoxide, potassium tert-butoxide, and lithium hexamethylamide;

and/or the second base is at least one of cesium carbonate, sodium tert-butoxide, potassium carbonate, potassium acetate and potassium phosphate;

and/or the third alkali is at least one of cesium carbonate, potassium carbonate, sodium hydroxide and lithium hydroxide.

As another preferable embodiment of the preparation method provided by the present invention, the catalyst is palladium tetratriphenylphosphine (Pd (PPh)₃)₄) Palladium acetate (Pd (OAc)₂) Tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (Pd) (dppf) Cl₂) At least one of;

and/or the brominating agent is at least one of N-bromosuccinimide, bromine and phenyl trimethyl ammonium tribromide.

The preparation method provided by the invention takes a compound II and ethyl formate as starting materials, and finally prepares the compound with the structure of the formula I by synthesizing an intermediate IV, an intermediate VI and an intermediate VII. The whole preparation process is simple to operate, easy to control, low in requirements on production equipment and suitable for industrial large-scale production.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound having the structure of formula i or a pharmaceutically acceptable salt thereof.

In the present invention, a compound having the structure of formula i or a pharmaceutically acceptable salt thereof is a pharmaceutically active ingredient in a pharmaceutical composition.

The present invention provides pharmaceutical compositions comprising, in addition to a therapeutically effective amount of at least one compound having the structure of formula i:

one or more pharmaceutical excipients;

and/or one or more pharmaceutically active substances having Polo-like kinase inhibitory activity other than the compound having the structure of formula I or a pharmaceutically acceptable salt thereof.

Methods of preparing the pharmaceutical compositions provided herein will be apparent to those skilled in the art and include conventional mixing, dissolving, lyophilizing and like techniques.

The pharmaceutical composition provided by the invention can be prepared into various common dosage forms, such as tablets, pills, capsules, granules, oral solutions, oral suspensions, oral emulsions, injections and the like, according to conventional preparation methods in the pharmaceutical field. It is convenient to provide the patient with clinical use by administering to the patient by various common modes of administration, such as oral or parenteral administration (by intravenous, intramuscular, topical or subcutaneous routes).

In a fourth aspect, the invention provides the use of a compound having the structure of formula i or a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising a compound having the structure of formula i or a pharmaceutically acceptable salt thereof as an inhibitor of PLK1 in the manufacture of a medicament for the treatment of tumours.

For the application of the compound and the pharmaceutical composition provided by the invention in preparing the medicine for treating the tumor, a mode of applying a therapeutically effective amount of the compound or the pharmaceutical composition provided by the invention to a patient needing to treat the tumor is adopted to realize the treatment effect. The compound or the pharmaceutical composition provided by the invention can be used alone as a treatment means for treating tumors, and can also be used in combination with other conventional treatment means for treating tumors, such as surgery, radiotherapy, chemotherapy and the like.

The therapeutically effective amount of the compounds or pharmaceutical compositions provided herein for treating tumors depends on a number of factors. May vary depending on the particular type of tumor that is to be treated, as can be determined by one skilled in the art without undue experimentation. The actual treatment will also take into account factors such as the age and weight of the patient, the severity of the condition, the particular mode of administration, etc., and will ultimately depend on the discretion of the attendant physician or clinician.

Further, the tumor is selected from:

skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, rectal cancer, esophageal cancer, tongue cancer, kidney cancer, cervical cancer, corpus uteri cancer, testicular cancer, urinary cancer, melanoma, astrocytoma, meningioma, hodgkin's lymphoma, non-hodgkin's lymphoma, acute lymphatic leukemia, chronic lymphatic leukemia, acute myeloid leukemia, chronic myeloid leukemia, adult T-cell leukemia lymphoma, hepatocellular carcinoma, multiple myeloma, basal cell tumor, seminoma, chondrosarcoma, myosarcoma, fibrosarcoma.

Some of the terms involved in the expression of the present invention are defined as follows:

the term "pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the parent compound. The salt comprises:

acid addition salts obtained by reaction of the free base of the parent compound with an inorganic acid or with an organic acid; the inorganic acid comprises hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid and the like; the organic acid includes acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, malonic acid, or the like;

or, a salt formed when an acid proton present in the parent compound is replaced with a metal ion or coordinated with an organic base; the metal ions include alkali metal ions, alkaline earth ions, aluminum ions and the like; the organic base includes ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.

The term "heteroaromatic ring" refers to an aromatic heterocyclic ring, typically a 5-to 8-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O or S. The aromatic heterocyclic ring may optionally be further fused or linked to aromatic and non-aromatic carbocyclic and heterocyclic rings. Non-limiting examples of such aromatic heterocycles include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, imidazolyl, thiazolyl, isothiazolyl, thiaoxazolyl, pyrrolyl, phenyl-pyrrolyl, furanyl, phenyl-furanyl, oxazolyl, isoxazolyl, pyrazolyl, thienyl, benzothienyl, isoindolyl, benzimidazolyl, indazolyl, quinolinyl, isoquinolinyl, 1, 2, 3-triazolyl, 1-phenyl-1, 2, 3-triazolyl, 2, 3-dihydroindolyl, 2, 3-dihydrobenzofuranyl, 2, 3-dihydrobenzothienyl, benzopyranyl, 2, 3-dihydrobenzoxazinyl, 2, 3-dihydroquinoxalinyl, and the like.

The term "pharmaceutical composition" refers to a mixture of one or more of the compounds provided herein with other chemical ingredients (e.g., pharmaceutical excipients). The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

The term "pharmaceutical excipient" refers to a substance that has been reasonably evaluated in terms of safety, does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the compound administered, and is included in a pharmaceutical preparation, in addition to an active ingredient. The pharmaceutic adjuvant has important functions of solubilization, dissolution assistance, sustained and controlled release and the like besides excipient, carrier and stability improvement, and is an important component which may influence the quality, safety and effectiveness of the pharmaceutical composition. The pharmaceutical excipients include, but are not limited to, carriers, diluents, excipients, solubilizers, binders, disintegrants, penetration enhancers, pH adjusters, buffers, release retardants, flavoring agents, preservatives, antioxidants, and the like.

The invention has the beneficial effects that:

(1) the invention provides a 4H-pyrano [2,3-c ] pyridine-4-ketone compound or a pharmaceutically acceptable salt thereof, which can be used as an effective PLK1 inhibitor and has stronger inhibitory activity.

(2) The invention also provides a pharmaceutical composition containing the 4H-pyrano [2,3-c ] pyridine-4-ketone compound or pharmaceutically acceptable salts thereof, and the pharmaceutical composition has good antitumor pharmacological activity.

Detailed Description

In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more apparent, the present invention is further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present patent and do not limit the scope of the invention in any way.

In the preparation method of the compound with the structure of formula I or the pharmaceutically acceptable salt thereof, the specific reaction process is as follows:

the first step is to synthesize intermediate III: dissolving a compound II, ethyl formate and a first alkali in a first solvent, and reacting at 0-120 ℃ to generate an intermediate III.

Wherein the first base is selected from at least one of sodium hydride, sodium tert-butoxide, potassium tert-butoxide, and lithium hexamethylamide.

The first solvent can be at least one selected from tetrahydrofuran, dioxane, toluene and acetonitrile.

For example, 1mol of compound II, 5mol of ethyl formate and 3mol of sodium tert-butoxide are dissolved in 2L of tetrahydrofuran and reacted with stirring at 60 ℃ to give intermediate III. The reaction can be monitored by Thin Layer Chromatography (abbreviated as TLC), and after the reaction is finished, the intermediate iii can be obtained by conventional treatment processes such as extraction, drying, concentration, separation and the like.

The second step is the synthesis of intermediate IV: and dissolving the intermediate III and a bromization reagent in a second solvent, and reacting at 0-80 ℃ to generate an intermediate IV.

Wherein, the brominating agent can be at least one of N-bromosuccinimide, bromine and phenyl trimethyl ammonium tribromide.

The second solvent may be at least one selected from tetrahydrofuran, dioxane, N-dimethylformamide, and acetonitrile.

For example, 1mol of intermediate III and 1.2mol of N-bromosuccinimide are dissolved in 2L of tetrahydrofuran and reacted with stirring at 40 ℃ to give intermediate IV. The reaction can be monitored by TLC, and the intermediate IV can be obtained by conventional treatment processes such as extraction, concentration, separation and the like after the reaction is finished.

The third step is the synthesis of intermediate VI: and dissolving the intermediate IV, the compound V, a catalyst and a second alkali in a third solvent, and reacting at 40-120 ℃ to generate an intermediate VI.

Wherein the catalyst can be selected from palladium tetratriphenylphosphine (Pd (PPh)₃)₄) Palladium acetate (Pd (OAc)₂) Tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) 1, 1' -bis (diphenylphosphino) ferrocene dichloropalladium (Pd) (dppf) Cl₂) At least one of (1).

The second base is at least one selected from cesium carbonate, sodium tert-butoxide, potassium carbonate, potassium acetate and potassium phosphate.

The third solvent may be any one selected from dioxane, N-dimethylformamide, toluene, N-dimethylacetamide, acetonitrile, dioxane/water.

For example, 1mol of intermediate IV, 1mol of compound V, 0.05mol of palladium tetratriphenylphosphine and 2mol of cesium carbonate are dissolved in 4L of acetonitrile and reacted with stirring at 80 ℃ to give intermediate VI. The reaction can be monitored by TLC, and the intermediate VI can be obtained by conventional treatment processes such as extraction, drying, concentration, separation and the like after the reaction is finished.

The fourth step is to synthesize an intermediate VII: and dissolving the intermediate VI and phosphorus oxychloride in a fourth solvent, and reacting at 20-120 ℃ to generate an intermediate VII.

Wherein, the fourth solvent can be at least one of acetonitrile, toluene and N, N-dimethylformamide.

For example, 1mol of intermediate VI and 2mol of phosphorus oxychloride are dissolved in 5L of toluene and reacted with stirring at 80 ℃ to give intermediate VII. The reaction can be monitored by TLC, and the intermediate VII can be obtained by conventional treatment processes such as extraction, drying, concentration, separation and the like after the reaction is finished.

The fifth step is the synthesis of a compound having the structure of formula i: and dissolving the intermediate VII, the compound VIII and a third alkali in a fifth solvent, and reacting at 20-120 ℃ to generate the compound with the structure of the formula I.

Wherein the third base is at least one of cesium carbonate, potassium carbonate, sodium hydroxide and lithium hydroxide.

The fifth solvent may be at least one selected from the group consisting of N, N-dimethylacetamide, N-dimethylformamide, toluene, acetonitrile, and dimethylsulfoxide.

For example, 1mol of intermediate VII, 1mol of compound VIII and 2mol of potassium carbonate are dissolved in 10L of toluene and reacted with stirring at 75 ℃ to give the compound having the structure of formula I. The reaction can be monitored by TLC, and after the reaction is finished, the final product can be obtained by conventional treatment processes such as extraction, drying, concentration, separation and the like.

The invention is described in more detail by referring to a part of the tests, which are carried out in sequence, and the following detailed description is given by combining specific examples:

unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the raw materials, instruments, equipment and the like used in the following examples are either commercially available or available by existing methods; the dosage of the reagent is the dosage of the reagent in the conventional experiment operation if no special description exists; the experimental methods are conventional methods unless otherwise specified.

Example 1

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1-phenylethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

The first step is as follows:

adding the compound 1a (30.6 g, 200.0 mmol), ethyl formate (74 g, 1.0 mol) and 60w% sodium hydride solution (24.0 g, 600.0 mmol) into toluene (500 mL) at room temperature, heating to 80 ℃ for reaction, detecting the reaction by TLC, cooling to room temperature after the reaction is finished, slowly adding saturated ammonium chloride solution (500 mL) for quenching the reaction, drying an organic layer, concentrating, and carrying out column chromatography separation to obtain 18.3g of off-white solid (intermediate 1 b), wherein the yield is 56.1%;

the second step is that:

dissolving the intermediate 1b (18.0 g, 110.4 mmol) in acetonitrile (200 mL), adding N-bromosuccinimide (NBS) (23.6 g, 132.5 mmol) at room temperature, stirring for reaction under constant temperature, detecting by TLC, adding water after the reaction to quench the reaction, extracting with ethyl acetate (300 mL × 2), concentrating the organic layer, and separating by column chromatography to obtain a light yellow solid (intermediate 1 c) 19.2g with a yield of 72.2%;

the third step:

intermediate 1c (12.1 g, 50.0 mmol), compound 1d (8.3 g, 50.0 mmol), palladium tetrakistriphenylphosphine ((Pd (PPh)₃)₄) (2.9 g, 2.5 mmol) and potassium carbonate (13.8 g, 100.0 mmol) are dissolved in dioxane (200 mL) and water (50 mL), the temperature is raised to 90 ℃, stirring reaction is carried out, after the reaction is finished, diluted hydrochloric acid is used for adjusting the pH to 5-6, ethyl acetate (300 mL multiplied by 2) is used for extraction, organic layers are combined, drying and filtering are carried out, column chromatography separation is carried out, 10.5g of white-like solid (intermediate 1 e) is obtained, and the yield is 74.2%;

the fourth step:

dissolving the intermediate 1e (10.0 g, 35.3 mmol) and phosphorus oxychloride (10.7 g, 70.7 mmol) in acetonitrile (400 mL), heating to reflux and stirring for reaction, monitoring the reaction by TLC, cooling to 0 ℃ after the reaction is finished, slowly adding a saturated sodium bicarbonate solution to quench the reaction, extracting an organic layer by using ethyl acetate (200 mL multiplied by 2), drying, filtering, concentrating, and carrying out column chromatography separation to obtain 7.4g of a white-like solid (intermediate 1 f), wherein the yield is 69.6%;

the fifth step:

intermediate 1f (301 mg, 1.0 mmol), compound 1g (121 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in N, N-Dimethylformamide (DMF) (50 mL), the reaction was stirred at 60 ℃, monitored by TLC, after completion of the reaction, the reaction was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 198mg of an off-white solid (compound 1), yield 51.3%, ESI (+) m/z = 387.1.

Example 2

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (pyridin-4-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 2a (122 mg, 1.0 mmol), cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), the temperature was raised to 60 ℃ and the reaction was stirred, followed by TLC monitoring, after completion of the reaction, water (50 mL) was added to quench the reaction, which was extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 226mg of an off-white solid (compound 2), yield 58.4%, ESI (+) m/z = 388.1.

Example 3

(S) -3- (1- ((3- (benzo [ d ] [1,3] dioxolan-5-yl) -4-oxo-4H-pyrano [2,3-c ] pyridin-6-yl) amino) ethyl) benzonitrile

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 3a (146 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give 246mg of off-white solid (compound 3) in 59.9% yield and ESI (+) m/z = 412.1.

Example 4

(S) -3- (1- ((3- (benzo [ d ] [1,3] dioxolan-5-yl) -4-oxo-4H-pyrano [2,3-c ] pyridin-6-yl) amino) ethyl) benzamide

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

the intermediate 1f (301 mg, 1.0 mmol), the compound 4a (164 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combining the organic layers, drying, concentration, and column chromatography to give 271mg of an off-white solid (compound 4), yield 63.2%, ESI (+) m/z = 430.1.

Example 5

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (4-chlorophenyl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 5a (155 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give an off-white solid (compound 5) 230mg, yield 54.8%, ESI (+) m/z = 421.1.

Example 6

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (3-fluorophenyl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 6a (139 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give 255mg of off-white solid (compound 6) with a yield of 63.1% and ESI (+) m/z = 405.1.

Example 7

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (2-methoxypyridin-4-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 7a (152 mg, 1.0 mmol), cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give 275mg of an off-white solid (compound 7) with a yield of 65.9% and ESI (+) m/z = 418.1.

Example 8

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (3-methoxyphenyl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 8a (151 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give an off-white solid (compound 8) 232mg, 55.8% yield, ESI (+) m/z = 417.1.

Example 9

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (4-chloro-3-fluorophenyl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 9a (173 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give 285mg of an off-white solid (compound 9) with a yield of 65.1% and ESI (+) m/z = 439.1.

Example 10

(S) -3- (benzo [ d ] [1,3] dioxolan-5-yl) -6- ((1- (6-methoxypyridin-3-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Synthesizing an intermediate 1f according to the methods of the first, second, third and fourth steps in example 1;

intermediate 1f (301 mg, 1.0 mmol), compound 10a (152 mg, 1.0 mmol), cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), the temperature was raised to 60 ℃ and the reaction was stirred, followed by TLC monitoring, after completion of the reaction, water (50 mL) was added to quench the reaction, which was extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 229mg of an off-white solid (compound 10), yield 54.9%, ESI (+) m/z = 418.1.

Example 11

(S) -3- (1H-indazol-6-yl) -6- ((1-phenylethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 1c was synthesized according to the first and second steps of example 1;

the first step is as follows:

intermediate 1c (12.1 g, 50.0 mmol), compound 11a (8.1 g, 50.0 mmol), 1' -bis (diphenylphosphino) ferrocene dichloropalladium (Pd (dppf) Cl₂) Dissolving (1.8 g, 2.5 mmol) and potassium carbonate (13.8 g, 100.0 mmol) in dioxane (200 mL) and water (50 mL), heating to 90 ℃, stirring for reaction, adjusting pH to 5-6 with dilute hydrochloric acid after reaction, extracting with ethyl acetate (300 mL multiplied by 2), combining organic layers, drying, filtering, and performing column chromatography to obtain 8.6g of off-white solid (intermediate 11 b) with the yield of 61.2%;

the second step is that:

dissolving the intermediate 11b (8.6 g, 30.8 mmol) and phosphorus oxychloride (14.0 g, 92.5 mmol) in acetonitrile (200 mL), heating to reflux and stirring for reaction, monitoring the reaction by TLC, cooling to 0 ℃ after the reaction is finished, slowly adding a saturated sodium bicarbonate solution to quench the reaction, extracting an organic layer by using ethyl acetate (200 mL multiplied by 2), drying, filtering, concentrating, and carrying out column chromatography separation to obtain 7.2g of a white-like solid (intermediate 11 c), wherein the yield is 78.7%;

the third step:

intermediate 11c (297 mg, 1.0 mmol), compound 1g (121 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, water (50 mL) was added to quench the reaction, extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 237mg of off-white solid (compound 11), yield 62.0%, ESI (+) m/z = 383.2.

Example 12

(S) -3- (1H-indazol-6-yl) -6- ((1- (pyridin-4-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 2a (122 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combination of organic layers, drying, concentration, and column chromatography to give 254mg of an off-white solid (compound 12) with a yield of 66.3% and ESI (+) m/z = 384.1.

Example 13

(S) -3- (1- ((3- (1H-indazol-6-yl) -4-oxo-4H-pyrano [2,3-c ] pyridin-6-yl) amino) ethyl) benzonitrile

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 3a (146 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, water (50 mL) was added to quench the reaction, extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 272mg of an off-white solid (compound 13) with a yield of 66.8% and ESI (+) m/z = 408.1.

Example 14

(S) -3- (1- ((3- (1H-indazol-6-yl) -4-oxo-4H-pyrano [2,3-c ] pyridin-6-yl) amino) ethyl) benzamide

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 4a (164 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give 266mg of an off-white solid (compound 14) with a yield of 62.6% and ESI (+) m/z = 426.2.

Example 15

(S) -6- ((1- (4-chlorophenyl) ethyl) amino) -3- (1H-indazol-6-yl) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 5a (155 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combination of organic layers, drying, concentration, and column chromatography to give 285mg of an off-white solid (compound 15) with a yield of 68.5% and ESI (+) m/z = 417.1.

Example 16

(S) -6- ((1- (3-fluorophenyl) ethyl) amino) -3- (1H-indazol-6-yl) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 6a (139 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combination of organic layers, drying, concentration, and column chromatography to give 302mg of an off-white solid (compound 16) with a yield of 75.5% and ESI (+) m/z = 401.1.

Example 17

(S) -3- (1H-indazol-6-yl) -6- ((1- (2-methoxypyridin-4-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 7a (152 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, water (50 mL) was added to quench the reaction, extracted with ethyl acetate (50 mL × 2), the organic layers were combined, dried, concentrated, and isolated by column chromatography to give 277mg of an off-white solid (compound 17) with a yield of 67.1% and ESI (+) m/z = 414.2.

Example 18

(S) -3- (1H-indazol-6-yl) -6- ((1- (3-methoxyphenyl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 8a (151 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combination of organic layers, drying, concentration, and column chromatography to give an off-white solid (compound 18) 268mg, yield 65.0%, ESI (+) m/z = 413.2.

Example 19

(S) -6- ((1- (4-chloro-3-fluorophenyl) ethyl) amino) -3- (1H-indazol-6-yl) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 9a (173 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), warmed to 60 ℃ and stirred for reaction, monitored by TLC for reaction, quenched after reaction with water (50 mL), extracted with ethyl acetate (50 mL × 2), the organic layers combined, dried, concentrated, and isolated by column chromatography to give an off-white solid (compound 19) 268mg, yield 61.8%, ESI (+) m/z = 435.1.

Example 20

(S) -3- (1H-indazol-6-yl) -6- ((1- (6-methoxypyridin-3-yl) ethyl) amino) -4H-pyrano [2,3-c ] pyridin-4-one

Intermediate 11c was synthesized according to the first and second steps of example 11;

intermediate 11c (297 mg, 1.0 mmol), compound 10a (152 mg, 1.0 mmol), and cesium carbonate (648 mg, 2.0 mmol) were dissolved in DMF (50 mL), heated to 60 ℃ and stirred for reaction, followed by TLC monitoring, after completion of the reaction, quenching with water (50 mL), extraction with ethyl acetate (50 mL × 2), combination of organic layers, drying, concentration, and column chromatography to give 255mg of an off-white solid (compound 20) with a yield of 61.7% and ESI (+) m/z = 414.2.

Biological evaluation

In the embodiment, the inhibition effect of the compounds 1-20 prepared in the embodiments 1-20 on PLK1 kinase is detected, and Rigoserinib is an internal control compound.

The experimental method comprises the following steps:

diluting the compound with DMSO to 10 concentration gradients for later use;

add 2.5. mu.L of compound solution to each well, and add 5. mu.L of a 2 Xsubstrate and 2 Xadenosine triphosphate (ATP) mixed solution. Simultaneously adding 5. mu.L of kinase buffer solution, 5. mu.L of 2 Xsubstrate and 2 XATP mixed solution as 0% phosphorylation control, adding 5. mu.L of kinase buffer solution, 5. mu.L of 2 Xphosphopeptide solution as 100% phosphorylation and 0% inhibition as controls, and incubating at room temperature for 1 hour; adding 5 mu L of PLK 1-containing kinase solution into each well, and shaking for 30 s; after incubation at room temperature for 1 hour, 5. mu.L of stop buffer was added to each well, and the reaction was stopped by shaking for 30 seconds. The wavelength of the excitation light is 400nm, reading the plate, and obtaining a compound IC by calculation₅₀。

The specific data are shown in the following table:

as can be seen from the table above, the compounds 1 to 20 all have certain inhibitory effects on PLK1 kinase, wherein the IC50 values of the compounds 2,3, 4, 9, 10, 14 to 17 and 19 to 20 on PLK1 kinase are less than 10 nM.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims (10)

1. A compound having the structure of formula I:

wherein R is₁Is a substituted or unsubstituted phenyl or aromatic heterocyclic group,

R₂is 1H-indazol-6-yl, benzo [ d][1,3]Dioxolan-5-yl.

2. A process for the preparation of a compound having the structure of formula i, or a pharmaceutically acceptable salt thereof, as claimed in claim 1, comprising the steps of:

and (3) synthesis of an intermediate III:

reacting a compound II with a structure shown in a formula II and ethyl formate in a first solvent in the presence of a first base at a first reaction temperature to generate an intermediate III with a structure shown in a formula III;

and (3) synthesizing an intermediate IV:

the intermediate III reacts in a second solvent under the action of a bromization reagent at a second reaction temperature to generate an intermediate IV with a structure shown in a formula IV;

synthesis of intermediate vi:

under the action of a catalyst, the intermediate IV and a compound V with a structure of a formula V are subjected to a coupling reaction in a third solvent in the presence of a second alkali at a third reaction temperature to generate an intermediate VI with a structure of a formula VI;

and (3) synthesizing an intermediate VII:

the intermediate VI reacts in a fourth solvent under the action of phosphorus oxychloride at a fourth reaction temperature to generate an intermediate VII with a structure shown in a formula VII;

synthesis of Compound I:

and reacting the intermediate VII and the compound VIII with the structure of the formula VIII in a fifth solvent in the presence of a third base at a fifth reaction temperature to generate the compound with the structure of the formula I.

3. The method according to claim 2, wherein the first solvent is at least one of tetrahydrofuran, dioxane, toluene, and acetonitrile;

and/or the second solvent is at least one of tetrahydrofuran, dioxane, N-dimethylformamide and acetonitrile;

and/or the third solvent is any one of dioxane, N-dimethylformamide, toluene, N-dimethylacetamide, acetonitrile and dioxane/water;

and/or the fourth solvent is at least one of acetonitrile, toluene and N, N-dimethylformamide;

and/or the fifth solvent is at least one of N, N-dimethylacetamide, N-dimethylformamide, toluene, acetonitrile and dimethyl sulfoxide.

4. The method of claim 2, wherein the first reaction temperature is 0 to 120 ℃;

and/or the second reaction temperature is 0-80 ℃;

and/or the third reaction temperature is 40-120 ℃;

and/or the fourth reaction temperature is 20-120 ℃;

and/or the fifth reaction temperature is 20-120 ℃.

5. The method of claim 2, wherein the first base is at least one of sodium hydride, sodium tert-butoxide, potassium tert-butoxide, lithium hexamethylamide;

and/or the second base is at least one of cesium carbonate, sodium tert-butoxide, potassium carbonate, potassium acetate and potassium phosphate;

and/or the third alkali is at least one of cesium carbonate, potassium carbonate, sodium hydroxide and lithium hydroxide.

6. The preparation method according to claim 2, wherein the catalyst is at least one of tetrakistriphenylphosphine palladium, palladium acetate, tris-dibenzylideneacetone dipalladium, 1' -bis (diphenylphosphino) ferrocene dichloropalladium;

and/or the brominating agent is at least one of N-bromosuccinimide, bromine and phenyl trimethyl ammonium tribromide.

7. A pharmaceutical composition comprising a compound having the structure of formula i as claimed in claim 1 or a pharmaceutically acceptable salt thereof.

8. Use of a compound having the structure of formula i according to claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 7 as an inhibitor of PLK1 in the manufacture of a medicament for the treatment of a tumour.

9. The use according to claim 8, wherein the tumor is selected from the group consisting of:

skin cancer, bladder cancer, ovarian cancer, breast cancer, stomach cancer, prostate cancer, colon cancer, lung cancer, bone cancer, brain cancer, rectal cancer, esophageal cancer, tongue cancer, kidney cancer, cervical cancer, corpus uteri cancer, testicular cancer, urinary cancer, melanoma, astrocytoma, meningioma, hodgkin's lymphoma, non-hodgkin's lymphoma, acute lymphatic leukemia, chronic lymphatic leukemia, acute myeloid leukemia, chronic myeloid leukemia, adult T-cell leukemia lymphoma, hepatocellular carcinoma, multiple myeloma, basal cell tumor, seminoma, chondrosarcoma, myosarcoma, fibrosarcoma.

10. The compound of claim 1 having the structure of formula i or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of compounds having the structure:

。